黑素再生中药对体外培养的黑素细胞酪氨酸酶基因和c-kit基因表达的影响

Effects of Melanocytes Regeneration Medicine on Cultured Melanocytes Tyrosinase Gene and c-kit Gene Expression

目的：分析黑素再生中药对体外培养的黑素细胞酪氨酸酶（Tyr）和c-kit基因表达的影响。方法：按照Halaban方法培养黑素细胞，传代至3～5代细胞用于实验；用黑素再生中药分别配制不同浓度的TICVA培养基。分散酶消化传代后第2天加入所配培养基，仔细观察细胞形态和生长状况，5d后计数；采用Bio-Rad蛋白分析试剂盒方法检测蛋白含量；蛋白质印记参照Halaban等方法。结果：高浓度黑素再生中药具有一定的细胞毒性，而8mg·mL-^1以下黑素再生液细胞毒性很小；黑素再生中药组细胞可溶性总蛋白质量浓度与对照组细胞可溶性总蛋白浓度，无显著差异（P＞0.05），但细胞可溶性总蛋白提高了6.7％；在等量的总蛋白中，黑素再生中药处理后的Tyr的含量增加了5.3％，黑素再生中药组c-kit表达增加了88.9％。结论：黑素再生中药治疗白癜风可能与Tyr基因的蛋白表达有一定的关联，对c-kit表达具有极显著的促进作用。

Objective： To study the effect of traditional Chinese medicine on the expression of melanocyte tyrosinase gene and c-kit in vitiligo. Methods：This was a vitro study Melanocytes Halaban accordance with the training methods, and passaged to 3 to 5 cells for experiments were used; Melanocytes renewable prepared Chinese medicines were different concentrations of TICVA medium. Scattered enzyme digestion after passage by the two days allocated to the medium, and carefully observe the growth of cell morphology, 5 days after counting. Protein content detected by Bio-Rad protein analysis kit. Protein imprint methods such as light Halaban. Results： Higher concentrations of black-renewable Chinese medicine has certain cytotoxicity, and 8 mg·mL-^1 following melanocytes renewable liquid cytotoxic smaller. The cells of the control group total soluble protein concentration and melanocyte cell regeneration medicine concentration of total soluble protein, no statistically significant difference was found （P〉O.05）, but the total soluble protein cells increased 6.7%; the contour of total protein, melanocytes renewable Chinese medicine treatment, the Tyr content increased 5.3%, black-renewable herbs on the expression of c-kit an increase of 88.9%. Conclusion： Melanocytes regeneration medicine and the treatment of vitiligo may Tyr protein gene expression has some association, expression of c-kit has a significant role in promoting.